Birth of viable female dogs produced by somatic cell nuclear transfer

Jang, Goo; Kim, M.K.; Oh, Hyun Ju; Hossein, Mohammad Shamim; Fibrianto, Yuda Heru; Hong, So Gun; Park, Jung E.; Kim, J.J.; Kim, H.J.; Kang, Sung Keun; Kim, Do-Yeop; Lee, B.C.

doi:10.1016/j.theriogenology.2006.11.006

Cited by 67 publications

(54 citation statements)

References 28 publications

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“…72(1): 89-92, 2010 Cloning technology using somatic cell nuclear transfer (SCNT) provides valuable opportunities for the fields of biomedical science, agriculture and preservation of endangered animals [2,6,17]. In canids, SCNT has been applied not only to the cloning of companion dogs [12,13,16] but also to the preservation of endangered canine species [14,18]. In some species, cloned offspring produced by SCNT have been reported to show normal reproductive characteristics and successfully produce progeny by natural mating [1,2,6,7,21].…”

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confidence: 99%

Serum Levels of Reproductive Hormones and Ultrasonographic Monitoring of Ovarian Follicles in Female Cloned Dogs

Hong

Park

et al. 2010

The Journal of Veterinary Medical Science

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ABSTRACT. This study investigated the reproductive characteristics in 2 cloned female beagle dogs (Clones A and B) during proestrus and early estrus. The pre-ovulatory estradiol peak occurred 2 days before the pre-ovulatory luteinizing hormone (LH) surge, while the follicle stimulating hormone surge started concomitantly with the LH surge in both cloned dogs. Serum progesterone levels increased during proestrus and estrus and its concentration on the day of the LH surge was 3.59 and 2.71 ng/ml in Clones A and B, respectively. Gradual follicular growth was observed by ultrasonography during proestrus. Although the numbers are limited, these cloned female dogs showed normal ranges of reproductive hormone levels and follicular changes during proestrous and early estrous stages of the cycle.KEY WORDS: cloned dog, estrous cycle, puberty, reproductive endocrinology/hormone, ultrasonography.J. Vet. Med. Sci. 72(1): 89-92, 2010 Cloning technology using somatic cell nuclear transfer (SCNT) provides valuable opportunities for the fields of biomedical science, agriculture and preservation of endangered animals [2,6,17]. In canids, SCNT has been applied not only to the cloning of companion dogs [12,13,16] but also to the preservation of endangered canine species [14,18]. In some species, cloned offspring produced by SCNT have been reported to show normal reproductive characteristics and successfully produce progeny by natural mating [1,2,6,7,21]. To our knowledge, however, there are no studies on the reproductive characteristics of cloned dogs. Accordingly, this study was designed to investigate systemic reproductive characteristics of cloned beagles, specifically their hormonal profiles and ovarian follicular development.Recently, we produced two cloned beagles (designated as Clones A and B) derived from canine fetal fibroblasts [10]. For this study, they were monitored regularly from six months of age for the presence of vulvar swelling and serosanguinous vaginal discharge, which were used as indications of the onset of proestrus. From the first day of proestrus, jugular venous blood samples were collected twice a day for hormonal analysis. Serum was prepared by centrifuging at 3,000 rpm for 20 min and stored at -30C until use for hormone measurement. Additionally, to gain more detailed information regarding the changes of hormones around the time of ovulation, blood samples were collected three times a day after the serum progesterone concentration exceeded 1 ng/ml. Previous studies demonstrated that progesterone can be used to accurately predict ovulation in canines [11,14].For analyzing reproductive hormones, progesterone, estradiol-17 (E 2 ), follicle stimulating hormone (FSH), and luteinizing hormone (LH), the assays were performed with the assistance of the Neodin Veterinary Laboratory, Seoul, South Korea (http://www.vetlab.co.kr). Briefly, serum progesterone concentrations were measured by radioimmunoassay (RIA) using a Coat-a-Count Progesterone Kit (Siemens Medical Solutions Diagnostics, U.S.A.). The kit cont...

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confidence: 99%

Serum Levels of Reproductive Hormones and Ultrasonographic Monitoring of Ovarian Follicles in Female Cloned Dogs

Hong

Park

et al. 2010

The Journal of Veterinary Medical Science

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“…Cumulus cells were removed and the oocytes were enucleated by micromanipulation (Jang et al, 2007). For the preparation of donor cells, 1 lg/ml of doxycycline was added to Tet-on eGFP cells the day before SCNT.…”

Section: Generation Of Doxycycline Inducible Transgenic Cloned Dogsmentioning

confidence: 99%

Generation of transgenic dogs that conditionally express green fluorescent protein

Kim

Park

et al. 2011

Genesis

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Summary: We report the creation of a transgenic dog that conditionally expresses eGFP (enhanced green fluorescent protein) under the regulation of doxycycline. Briefly, fetal fibroblasts infected with a Tet-on eGFP vector were used for somatic cell nuclear transfer. Subsequently reconstructed oocytes were transferred to recipients. Three clones having transgenes were born and one dog was alive. The dog showed all features of inducible expression of eGFP upon doxycycline administration, and successful breeding resulted in eGFPpositive puppies, confirming stable insertion of the transgene into the genome. This inducible dog model will be useful for a variety of medical research studies. genesis 49:472-478, 2011. V V C 2011 Wiley-Liss, Inc.

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“…Quatre chiots, appartenant à deux portées, ont été obtenus à partir d'embryons issus du transfert de cellules cutanées auriculaires adultes (Lee et al 2005 ;Parker et al 2006 ;Jang et al 2006). Les ovocytes ont été maturés in vivo, collectés par lavage des oviductes.…”

Section: Clonageunclassified

Biotechnologies de l'embryon dans l'espèce canine

Chastant‐Maillard¹,

Saint‐Dizier²,

Lesegno³

et al. 2007

bavf

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La biologie de l'ovocyte et de l'embryon canins, qui présentent des particularités spécifiques, est très mal connue. La chienne se distingue principalement par les modalités de reprise de la méiose ovocytaire : celle-ci n'a pas lieu au moment de l'ovulation, comme chez les autres femelles mammifères, mais dans l'oviducte 48 à 60 heures après l'ovulation. Les facteurs responsables de ce retard ne sont pas connus. Ceci explique sans doute pourquoi les taux de maturation in vitro obtenus à l'heure actuelle sont faibles (10 à 30 %). La collecte des ovocytes est aussi un facteur limitant majeur, en l'absence de protocole efficace d'induction des cycles et de superovulation. Les rendements de fécondation in vitro sont également faibles (10 %), avec un taux élevé de polyspermie. À l'heure actuelle, aucun chiot n'est encore né à partir d'un embryon produit in vitro. Quant aux embryons produits in vivo, leur collecte est difficile pour des raisons anatomiques et le rendement est limité par l'impossibilité d'induire des superovulations ; par ailleurs, leur transfert chez des femelles receveuses se heurte aux difficultés de synchronisation des ovulations entre la femelle donneuse et les receveuses, et la littérature ne décrit que six essais, qui ont abouti à la naissance de 45 chiots. Avec un très faible recours à la culture in vitro, quatre naissances de chiots ont été obtenues par clonage de cellules somatiques. Les biotechnologies de la reproduction sont donc largement en retard dans l'espèce canine, qui souffre d'un manque de travaux fondamentaux visant à mieux comprendre ses mécanismes physiologiques spécifiques. Ce déficit est d'autant plus dommageable que le chien prend une place croissante et pertinente en tant que modèle biomédical. Mots

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Birth of viable female dogs produced by somatic cell nuclear transfer

Cited by 67 publications

References 28 publications

Serum Levels of Reproductive Hormones and Ultrasonographic Monitoring of Ovarian Follicles in Female Cloned Dogs

Serum Levels of Reproductive Hormones and Ultrasonographic Monitoring of Ovarian Follicles in Female Cloned Dogs

Generation of transgenic dogs that conditionally express green fluorescent protein

Biotechnologies de l'embryon dans l'espèce canine

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